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18β-Glycyrrhetinic acid altered the intestinal permeability in the human Caco-2 monolayer cell model

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European Journal of Nutrition Aims and scope Submit manuscript

A Correction to this article was published on 21 February 2023

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Abstract

Purpose

Glycyrrhizin (GL) and its metabolites 18α-glycyrrhetinic acid (18α-GA) and 18β-glycyrrhetinic acid (18β-GA) are used as traditional medicine and food sweeteners. As the major rout of their administration is oral way, therefore their impact on intestinal epithelial cells are investigated.

Methods

The effects of GL and its metabolites on cell viability using MTT assay, on cytotoxicity using LDH release, on integrity of intestinal epithelial cells by measuring the transepithelial electrical resistance (TEER) and Luciferase permeability tests, on the expression of tight junction proteins at mRNA and protein level by qPCR and western blot techniques, and ultimately on the rate of test compounds absorption via Caco-2 cells monolayer were investigated.

Results

MTT assay showed a concentration- and time-dependent decrease in metabolic activity of Caco-2 cells induced by GL, 18α-GA, and 18β-GA, while only 18β-GA increased the LDH leakage. The monolayer integrity of Caco-2 cells in TEER assay only was affected by 18β-GA. The permeability of paracellular transport marker was increased by 18α-GA and 18β-GA and not GL. In transport studies, only metabolites were able to cross from Caco-2 cells monolayer. qPCR analyses revealed that 18β-GA upregulated the expression of claudin-1 and -4, occludin, junctional adhesion molecules and zonula occludens-1, while 18α-GA upregulated only claudin-4. The expression of claudin-4 at protein level was downregulated non-significantly at 50 μM concentration of 18β-GA.

Conclusion

Our results suggest that 18β-GA may cause cellular damages at higher concentrations on gastrointestinal cells and requires a remarkable attention of the nutraceutical and pharmaceutical industries.

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All data generated or analyzed during this study are included in this published article.

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Acknowledgements

The authors would like to thank Professor Johanna Fink-Gremmels and Dr. Peyman Akbari from Utrecht University, for their constructive comments.

Funding

This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

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Authors and Affiliations

  1. Experimental and Applied Pharmaceutical Sciences Research Center, Urmia University of Medical Sciences, Urmia, Iran

    Mojtaba Malekinejad & Hassan Malekinejad

  2. Department of Internal Medicine, School of Medicine, Urmia University of Medical Sciences, Urmia, Iran

    Mojtaba Malekinejad & Mohammad Reza Pashaee

  3. Department of Pharmacology and Toxicology, School of Pharmacy, Urmia University of Medical Sciences, Urmia, Iran

    Hassan Malekinejad

Authors
  1. Mojtaba Malekinejad
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  2. Mohammad Reza Pashaee
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Contributions

MM carried out the experiments and wrote the first draft of the manuscript. MRP designed the study and participated in data collection and analyses. HM carried out experiments, participated in the statistical analyses and in the drafting of manuscript and discussed the results. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Mohammad Reza Pashaee or Hassan Malekinejad.

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Malekinejad, M., Pashaee, M.R. & Malekinejad, H. 18β-Glycyrrhetinic acid altered the intestinal permeability in the human Caco-2 monolayer cell model. Eur J Nutr 61, 3437–3447 (2022). https://doi.org/10.1007/s00394-022-02900-4

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  • DOI: https://doi.org/10.1007/s00394-022-02900-4

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